Fig. 3

Increased Tlr7 expression in B cells from pristane-treated mice. Spleen cells from untreated mice were stained with anti-CD19, CD138, IgM, and IgD antibodies and four B cell subsets were identified by flow cytometry and sorted. a Gating strategy for isolating naïve B cells (CD19⁺CD138⁻IgM⁺IgD⁺, NB), switched memory B cells: CD19⁺CD138⁻IgM⁻IgD⁻, sMB), switched plasmablasts (CD19⁺CD138⁺IgM⁻IgD⁻, sPB) and plasma cells (CD19^+/−CD138⁺⁺IgM⁻IgD⁻, PC). b Top, CIITA gene expression (Q-PCR) (compared to 18S rRNA) in PC, sPB, sMB, and NB subsets. Bottom, spleen cells were stained with anti-CD19, CD138, IgM, IgD, and major histocompatibility complex (MHC) II and analyzed by flow cytometry. Mean fluorescence intensity (MFI) of MHC II staining in these four B cell subsets is indicated. c Tlr7 gene expression (compared to 18S) in purified sMB, sPB, PC, and NB subsets from pristane-treated and PBS-treated mice. d IgG production stimulated by R848 in sPB + PC isolated from pristane or PBS-treated mice and cultured with R848 or PBS. IgG secretion in culture supernatants was measured at 10 days (ELISA). e IgG production stimulated by R848 in sMB. f Tlr7 gene expression (Q-PCR) (compared to 18S) in magnetic bead-purified splenic CD19⁺ B cells from PBS-treated mice stimulated by PBS, interferon (IFN)α (80U/ml), R848 (1 μg/ml) or IFNα (80U/ml) plus R848 (1 μg/ml) for 6 h: *p <0.05, **p <0.01, Mann–Whitney test